Myosin light chain isoforms and their phosphorylation in arterial smooth muscle

F. Erdődi, M. Barany, K. Barany

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

Arterial smooth muscle myosin contains nonphosphorylated and phosphorylated light chains that appear as 4 spots on two-dimensional, Coomassie blue-stained gel electrophoretograms at the 20,000-molecular weight level (referred to as spots 4 through 1 in order of decreasing isoelectric points). Anti-light chain recognizes the proteins in all 4 light chain spots. Complete dephosphorylation of light chain in muscle homogenate, by inhibiting myosin light chain kinase and by adding phosphatase, leads to 2 spots on two-dimensional gel electrophoretograms; both spots are visible on immunoblots. Stimulation (K+ or stretch) of smooth muscle results in increased light chain phosphorylation. Autoradiography of the gel electrophoretograms reveals that radioactive components are contained in spots 3, 2, 1, and in an additional spot with lower isoelectric point, referred to as spot 0. Phosphoamino acid analysis shows that spots 3 and 1 contain phosphoserine, whereas spots 2 and 0 contain phosphoserine and phosphothreonine. Two-dimensional phosphopeptide mapping of the trypsin-digested proteins from spots 3 and 1 shows predominantly 2 peptides; whereas from spots 2 and 0, it shows 5 peptides. Sodium dodecyl sulfate gel electrophoresis of the phosphopeptides obtained with Staphylococcus aureus V8 digestion gives identical maps for spots 3 and 2, which are different from the identical maps of spots 1 and 0. The results suggest that arterial smooth muscle myosin contains 2 nonphosphorylated 20,000-dalton light chain isoforms with different amino acid sequences and that each isoform can be mono- and diphosphorylated.

Original languageEnglish
Pages (from-to)898-903
Number of pages6
JournalCirculation Research
Volume61
Issue number6
Publication statusPublished - 1987

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Myosin Light Chains
Smooth Muscle
Protein Isoforms
Phosphorylation
Light
Gels
Smooth Muscle Myosins
Phosphoserine
Phosphopeptides
Isoelectric Point
Phosphothreonine
Phosphoamino Acids
Myosin-Light-Chain Kinase
Peptides
Autoradiography
Phosphoric Monoester Hydrolases
Sodium Dodecyl Sulfate
Trypsin
Staphylococcus aureus
Electrophoresis

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

Myosin light chain isoforms and their phosphorylation in arterial smooth muscle. / Erdődi, F.; Barany, M.; Barany, K.

In: Circulation Research, Vol. 61, No. 6, 1987, p. 898-903.

Research output: Contribution to journalArticle

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